Reconstitution of G Protein-Coupled Receptors into a Model Bilayer System: Reconstituted High-Density Lipoprotein Particles
Reconstituted high-density lipoprotein particles (rHDL) are powerful platforms used as a model phospholipid bilayer system to study membrane proteins. They consist of a discoidal-shaped planar bilayer of phospholipids that is surrounded by a dimer of apolipoprotein A-I (apoA-I). The amphipathic nature of apoA-1 shields the hydrophobic acyl chains of the lipids from solvent and keeps the particles soluble in aqueous environments. These monodispersed, nanoscale discoidal HDL particles are approximately 10–11 nm in diameter with a thickness that is dependent on the length of the phospholipid acyl chain. Reconstituted HDL particles can be assembled in vitro using purified apoA-1 and purified lipids. Investigators have utilized this model bilayer system to co-reconstitute membrane proteins, and take advantage of the small size and its monodispersion. Our laboratory and others have utilized the rHDL approach to study the behavior of G protein-coupled receptors. In this chapter, we describe strategies for the preparation of rHDL particles containing GPCRs in their monomeric form and discuss various methodologies used to analyze the reconstituted receptor function.
Key wordsApolipoprotein A-I High-density lipoprotein particles Receptor 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine 1-Palmitoyl-2-oleoyl-sn-glycero-3-[phosphor-rac-(1-glycerol)] Monomer Oligomer
This work is supported through funding of the National Institutes of Health (GM-068603 and GM-083118), the University of Michigan Biological Sciences Scholars Program and the Cellular and Molecular Biology Training Grant and the University of Michigan Rackham Merit Program.
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